福建省稻田土壤细菌群落的16S rDNA-PCR-DGGE分析

Analysis of the Bacterial Communities in Paddy Soils in Fujian Province Using 16S rDNA -PCR- DGGE

摘要:

用不依赖细菌培养的16S rDNA-PCR-DGGE方法对福建省6个不同地区12个取样点的稻田土壤进行细菌群落结构分析。对12份样品直接提取其总DNA, 用F341GC/R534引物扩增16S rDNA基因的 V3可变区, 结合DGGE(denaturing gradient gel electrophoresis)技术分析样品细菌群落组成。结果表明, 福建省不同地区的稻田土壤之间细菌群落结构存在较大差异, 大体上可分为闽东、闽南、闽北、闽西4个大类。同一地区的根际土和表土样品之间也存在差异, 但差异相对较低, 其中龙岩根际土和表土细菌群落结构相似性最大, 永泰差异性最大。回收了DGGE图谱中11个条带, 测序结果经过Blast比对表明其中10个条带代表的细菌是不可培养的, 显示了DGGE技术的优越性。

Abstract:

16S rDNA-PCR-DGGE, a cultivation-independent approach, was used to analyze the bacterial communities in paddy soils in Fujian Province. The bulk soils and rhizosphric soils were sampled from six different ecological soil regions. Total DNA was directly extracted and amplified with the F341GC and R534 primers targeting the 16S rDNA V3. The amplified fragments were analyzed by perpendicular DGGE. Cluster analysis revealed that there was a high diversity of bacterial community compositions among different soil samples tested. Basically they could be grouped to Mindong (East), Minnan (South), Minbei (North) and Minxi (West) ecological regions. 16S rDNA-PCR-DGGE profiles from bulk and rhizospheric soil in the same region showed less bacterial diversity but more similarity. The bacterial community from bulk soil and rhizospheric soil in Longyan shared the most similar DGGE banding patterns, and the banding patterns in Yongtai exhibited the highest diversity. Eleven DGGE bands recovered were re-amplified, sequenced and aligned with Blast. The results indicated that ten of the fragments belong to uncultivated bacteria implying DGGE technique having priority in analyzing uncultivated bacteria in the paddy soils.